Suspension compositions of physiologically active phenolic compounds and methods of making and using the same

ABSTRACT

The present invention is directed to compositions comprising physiologically active phenolic compounds and methods for making and using the same. In particular embodiments, the compositions described herein include suspension formulations including a physiologically active phenolic compound provided as a nanoparticulate material and dispersed within an edible lipid.

TECHNICAL FIELD

The present disclosure relates to compositions including physiologicallyactive phenolic compounds and methods for producing and utilizing suchcompositions. In certain embodiments, the compositions described hereincomprise a nanoparticulate phenolic compound dispersed in an ediblelipid, and in further embodiments, methods of making and using suchcompositions are provided.

BACKGROUND

There are a variety of physiologically active phenolic substances,including naturally-occurring phenolic compounds, that have been shownto exhibit favorable medicinal or nutritional properties. However, for avariety of reasons, it is often difficult to provide these phenoliccompounds in a composition that is readily ingested and suited fordelivering the compounds to subjects in sufficient amounts to achieve adesired nutritional or therapeutic effect.

Many phenolic substances are not readily bioavailable. As a result, inorder to achieve any benefit from consumption or administration of thephenolic substance, a subject must consume or be administered largequantities of the phenolic compound. In addition, oral delivery of manydesirable phenolic compounds has proven difficult or unpalatable tosubjects, which can lead to a perceived requirement for parenteralroutes of administration and/or the formulation of relatively complex,multi-component, pharmaceutical grade compositions designed to increasethe targeted compound's bioavailability.

DESCRIPTION OF THE FIGURES

FIGS. 1 and 2 show the particle size distribution of two differentgrades of an exemplary physiologically active phenolic compound prior topreparation according to the methods described herein or inclusion in acomposition according to the present description. FIG. 1 provides theparticle size distribution of Genivida™ (food grade genistein) uponreceipt from a commercial supplier. FIG. 2 provides the particle sizedistribution of Bonistein® (pharmaceutical grade genistein) upon receiptfrom a commercial supplier.

FIGS. 3A-3C show the initial and 3-month particle size distributions fortwo different grades of an exemplary nanoparticulate, physiologicallyactive, phenolic compound material (nanoparticulate genistein) preparedaccording to the methods described herein and included withincompositions according to the present description.

FIG. 4 provides particle size distribution data for an exemplaryphysiologically active phenolic compound (genistein) prepared accordingto the present description and included within a composition asdescribed herein. The genistein was sourced in two different grades, apharmaceutical grade material (Bonistein®) and a food grade material(Genivida™). FIG. 4 provides initial particle size distribution data forthe genistein materials prepared according to the methods describedherein and included in a composition according to the presentdescription (“Initial Measurement”). FIG. 4 also provides particle sizedistribution data for the same genistein materials after storage forthree months (“3-Month Measurement”) at ambient temperature.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

Compositions of physiologically active phenolic compounds are describedherein. In certain embodiments, the compositions described herein aresuitable for administration to or consumption by a subject as apharmaceutical formulation, a medical food, or a dietary supplement. Inparticular embodiments, the compositions described herein include aphenolic compound provided as a nanoparticulate material suspendedwithin an edible lipid. Providing the phenolic compound asnanoparticulate material serves to increase the bioavailability of thephenolic compound and may additionally facilitate administration orconsumption of the phenolic compound at amounts sufficient to achieve adesired nutritional or therapeutic benefit.

Compositions prepared according to the present description have beenshown to exhibit desirable physical stability characteristics. Often,where active compounds are provided as a nanoparticulate material,particle agglomeration over time causes a significant shift in theparticle size distribution (PSD) of the active material, resulting in aloss of the advantages sought by providing the active as a nano-sizedmaterial in the first place. Embodiments of the compositions describedherein serve to preserve the PSD of the phenolic compound includedtherein over time so that, even after months of storage, the PSD of thephenolic compound is maintained as a nanoparticulate material.

Methods for preparing the compositions described herein are alsoprovided. In general, a physiologically active phenolic compound isprovided as a nanoparticulate material and then dispersed within anedible lipid, resulting in a suspension suitable for administration toor consumption by a subject. Any suitable size reduction process may beutilized to reduce the phenolic compound to an active materialexhibiting a desired, nano-sized PSD. In certain embodiments, thenanoparticulate phenolic compound material may be provided by anano-milling procedure. The term “active material,” as it is used in thepresent disclosure, refers to an amount of one or more physiologicallyactive phenolic compound(s).

Nano-mills typically include a milling media (e.g., balls, beads,pellets, satellites, crystalline media, etc.) into which the activematerial to be size-reduced is introduced. Once the active material isintroduced into the milling media, the milling media is agitated and theactive material is subjected to grinding and shearing forces that reducethe material's PSD. In order to facilitate milling, the active materialto be size-reduced can be provided in a liquid vehicle or a liquidvehicle may be introduced into the milling media. In specificembodiments of the methods described herein, the vehicle used innano-milling the active material may be an edible lipid suitable for usein a composition according to the present description.

The compositions described herein are suitable for consumption by oradministration to human and/or animal subjects. A composition accordingto the present disclosure may be prepared as a food-grade composition.Alternatively, a composition as described herein may be prepared aspharmaceutical-grade composition. In certain embodiments, thecompositions described herein may be prepared for direct administrationor consumption. In other embodiments, the compositions described hereinmay be prepared for combination with one or more other constituentsprior to administration or consumption.

I. Definitions

As used herein and in the appended claims, the singular forms “a,” “an,”and “the” include plural reference unless the context clearly dictatesotherwise.

Ranges can be expressed herein as from “about” one particular value,and/or to “about” another particular value. When such a range isexpressed, another embodiment includes from the one particular valueand/or to the other particular value. Similarly, when values areexpressed as approximations, by use of the antecedent “about,” it willbe understood that the particular value forms another embodiment. Itwill be further understood that the endpoints of each of the ranges aresignificant both in relation to the other endpoint, and independently ofthe other endpoint. It is also understood that there are a number ofvalues disclosed herein, and that each value is also herein disclosed as“about” that particular value in addition to the value itself. Forexample, if the value “10” is disclosed, then “about 10” is alsodisclosed. It is also understood that each unit between two particularunits are also disclosed. For example, if 10 and 15 are disclosed, then11, 12, 13, and 14 are also disclosed.

As used herein, “nanoparticulate” refers to material exhibiting a volumediameter, as measured using laser light diffraction, wherein the D(0.50) of the material is 0.5 μm or less. In certain embodiments, theterm “nanoparticulate” refers to material exhibiting a volume diameter,as measured using laser light diffraction, wherein the D (0.50) of thematerial is 0.5 μm or less and the D (0.90) is 2 μm or less. Particlesize analysis using laser light diffraction is a technique based onlight being scattered through various angles which are directly relatedto the size of the particles. By measuring the angles of light scatteredby the particles being analyzed and the intensity of this scatteredlight, a particle size distribution can be calculated. Techniques foruse in analyzing particle size in the context of the present disclosurecan be referred to as static light scattering, Rayleigh lightscattering, low angle light scattering (LALS), multiple angle lightscattering (MALS) Fraunhofer diffraction, or Mie Scattering. Measurementof particle size distributions using Mie Scattering allows for thedetermination of particle size distributions through the directionmeasurement of mass.

Two theoretical applications to the analysis of particle size by laserlight diffraction are based on assumptions about the properties of theparticles. Fraunhofer theory considers the following: particles arespherical, non-porous and opaque; particle diameters are greater thanthe wavelength of the laser light used in the analysis; and particlesare distant enough from each other not to interfere in the diffractionof light, exhibit random motion, and diffract light with the sameefficiency regardless of size and shape. Mie theory considers thedifferences in refractive index between the particles and the suspendingmedium, which allows the measurement technique to account for particlesin the size range below the wavelength of the laser light used in theanalysis. The relative amounts of different size particles aredetermined by measuring the intensity of light scattered at differentangles. As the particles get close to or smaller than the wavelength oflight, more of the light intensity is scattered to higher angles andback-scattered. Mie Scattering Theory accounts for this differentbehavior. In order to make particle size measurements, the lightintensity pattern is measured over the full angular range. When theparticle size is larger than the wavelength of the incident light, theMie equation reduces to the Fraunhofer equation. An array of detectors,including high-angle and back-scatter detectors, and multiple lightsources of different wavelengths are typically employed to allowmeasurement of the full size range in one analysis. Equipment suited foruse in analyzing particle size by laser light diffraction iscommercially available and manufactured, for example, by HoribaInstruments, Irvine, Calif.

In the context of the present description, the particle sizedistribution of a given material is provided in volume diameter asmeasured in accordance with USP 429 using a laser diffraction particlesize analyzer operating in the Mie Scattering Theory diffraction modeand equipped with a suspension dispersion sample chamber (e.g., asavailable from Horiba Instruments, Irvine, Calif., USA). For purposes ofthe present description, volume diameter is given as a particle sizedistribution defined by one or more of D (0.10), D (0.50) and D (0.90).When referred to herein, the term D (0.10) indicates the volumefrequency distribution of particles for which 10% of the sample is belowthe referenced size, the term D (0.50) indicates the volume frequencydistribution of particles for which 50% of the sample is below thereferenced size, and the term D (0.90) indicates the volume frequencydistribution of particles for which 90% of the sample is below thereferenced size. In addition to measurement of the particle sizedistribution via laser diffraction, the results from such particle sizeanalysis and/or the morphology of the particles may be confirmed usingknown electron microscopy techniques.

A “subject” for purposes of this disclosure is an animal to which acomposition as described herein can be administered in order to achievea targeted benefit. In certain embodiments, the subject is a humanbeing.

II. Compositions of Physiologically Active Phenolic Compounds

The compositions described herein include one or more physiologicallyactive phenolic compound. The phenolic compound may be a non-naturallyoccurring or a naturally occurring compound. In addition, the phenoliccompound may be chemically synthesized (even if also produced by naturalsources) or isolated, purified, or derived from natural sources. Inparticular embodiments, the phenolic compound is a naturally occurringcompound found in or derived from one or more species or varieties ofplant. Examples of such compounds suitable for use in the compositionsdescribed herein include, but are not limited to, compounds selectedfrom isoflavones, curcuminoids, flavonols, and stilbenoids.

Curcuminoids are natural phenols found, for example, in turmeric.Curcuminoids include curcumin, desmethoxycurcumin, andbis-desmethoxycurcumin. Curcumin can exist in several tautomeric forms,including a 1,3-diketo form and two equivalent enol forms. Curcuminoidsgenerally exhibit poor water solubility and poor bioavailability. Thechemical name for curcumin is(1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione(IUPAC). The chemical name for desmethoxycurcumin is(1E,6E)-1,6-Heptadiene-3,5-dione,1-(4-hydroxy-3-methoxyphenyl)-7-(4-hydroxyphenyl) (IUPAC). The chemicalname for bis-desmethoxycurcumin is (1E,6E)-1,7-bis(4-hydroxyphenyl)hepta-1,6-diene-3,5-dione (IUPAC). Curcuminoids are commerciallyavailable and can be isolated from plant material or manufactured inpurified form using known chemical syntheses.

Isoflavones include a variety of naturally occurring compounds andbelong to a class of organic compounds related to isoflavonoids. Manyisoflavones act as phytoestrogens in mammals, and some are termedantioxidants because of their ability to trap singlet oxygen.Isoflavones are produced almost exclusively by the members of theFabaceae family (i.e., the Leguminosae or bean family). Soybeans are themost common source of isoflavones in human food, and the majorisoflavones in soybean are genistein and daidzein. In general,isoflavones also exhibit poor water solubility and bioavailability.Genistein is commercially available and may be obtained in synthetic,purified form. Synthetic genistein is available, for example, asBONISTEIN from DSM Nutritional Products (DSM Nutritional Products, Inc.Parsippany, N.J.). Genistein's chemical name is5,7-dihydroxy-3-(4-hydroxyphenyl)-chromen-4-one (IUPAC). Daidzein'schemical name is 7-hydroxy-3-(4-hydroxyphenyl) chromen-4-one (IUPAC).Like genistein, daidzein is commercially available and can be extractedor isolated from plant material or manufactured in purified form usingknown chemical syntheses.

Flavanols that may be utilized in compositions according to the presentdescription include catechins. As used herein, the term “catechin”refers to a family of compounds as well as the compound itself. Catechinhas the chemical name(2R,3S)-2-(3,4-dihydroxyphenyl)-3,4-dihydro-2H-chromene-3,5,7-triol(IUPAC). The family of catechin compounds includes, for example,epicatechin, and epigallocatechin gallate (EGCG), which may also be usedin compositions, described herein. As used herein, “epicatechin” refersto one or both of two isomers of catechin in the cis configuration, andwhen used herein as a compound name, the term “catechin” refers to oneor both of two isomers of catechin in the trans configuration. Thechemical name for EGCG is[(2R,3R)-5,7-dihydroxy-2-(3,4,5-trihydroxyphenyl)chroman-3-yl]3,4,5-trihydroxybenzoate(IUPAC). Catechins are commercially available and can be found in andisolated from, for example, green tea, cocoa beans, the kola nut, rawapples, apricots, nectarines, pears and plums, blackberries, redraspberries, cranberries, cherries, broad beans. Catechins can also bemanufactured in purified form using known chemical syntheses.

Stilbenoids are naturally occurring phenols produced by severaldifferent plants. A stilbenoid of particular interest in the context ofthe compositions described herein is resveratrol. The chemical name forresveratrol is 5-[(E)-2-(4-hydroxyphenyl)ethenyl]benzene-1,3-diol(IUPAC). Like other stilbenoids, resveratrol produced naturally byseveral different species of plants, and it is often extracted forcommercial purposes from the roots of the Japanese Knotweed. Like theother phenol compounds specifically described herein, resveratrol iscommercially available and can be isolated from naturally occurringplant material or manufactured in purified form using known chemicalsyntheses.

The composition described herein may include one or more phenoliccompounds as described herein. For example, in particular embodiments,the compositions include a phenolic compound selected from curcumin,desmethoxycurcumin, bis-desmethoxycurcumin, genistein, daidzein,catechin, epicatechin, EGCG, resveratrol, and combinations of two ofmore of such compounds. In each embodiment, one or more phenoliccompound included in the composition is provided as a nanoparticulatematerial. In specific embodiments, the compositions disclosed herein mayinclude one or more phenolic compounds provided as nanoparticulatematerial exhibiting a D (0.50) of 0.4 μm or less. In certain suchembodiments, the compositions disclosed herein may include one or morephenolic compounds provided as nanoparticulate material exhibiting a D(0.50) selected from a D (0.50) of 0.35 μm or less, a D (0.50) of 0.30μm or less, a D (0.50) of 0.25 μm or less, and a D (0.50) of 0.20 μm orless. In addition to being characterized by a D (0.50) as detailedherein, the nanoparticulate active material may exhibit a D (0.90) of2.0 μm or less. In particular embodiments, the nanoparticulate activematerial exhibits a D (0.50) as detailed herein and a D (0.90) selectedfrom a D (0.90) of 1.5 μm or less, a D (0.90) of 1.0 μm or less, and a D(0.90) of 0.5 μm or less.

Nanoparticulate active material suitable for use in the compositionsdisclosed herein may be prepared according to known methods forproducing materials exhibiting a sub-micron PSD. In one embodiment,naturally derived or synthetically manufactured phenolic compoundmaterial may be nanomilled according to milling techniques known in theart. Nanomilling may include wet bead milling utilizing an agitator beadmill in a grinding container for continuous dispersion and fine wetgrinding. Alternatively, the necessary energy for dispersion andgrinding of a material within the milling media of a nanomill may betransmitted to the grinding media through agitator discs mounted on anagitator shaft. Several different nanomills and milling techniques arecommercially accessible and offered from, for example, CB Mills, ofGurnee, Ill., and NETZSCH Premier Technologies, LLC of Exton, Pa.

Though nanomilling is generally referenced herein as a means forproducing nanoparticulate active material suitable for use in thecompositions described herein, the nanoparticulate active material canbe produced by other suitable techniques as well. For example, thedesired phenolic compound may be provided as a nanoparticulate materialthrough one or more known wet milling techniques, super-critical orcompressed fluid techniques, hot or high-pressure homogenization,emulsification techniques, evaporative precipitation, antisolventprecipitation, microprecipitation, cryogenic techniques, complexationtechniques, ultrasonication techniques, solid dispersion techniques, orspray drying and lyophilization techniques.

The active material included in the compositions described herein isdispersed within an edible lipid to form a nanoparticulate suspension ofthe phenolic compound. The term “edible lipid” refers to naturallyoccurring and synthetic lipids, fats, and oils suitable for consumptionby or administration to a subject. In certain embodiments, the ediblelipid may be an edible glyceride selected from glycerides having a chainlength ranging from one carbon acetate to 22 carbons. Where an edibleglyceride is used as the edible lipid, the fatty acid chain of theedible glyceride may be saturated or exhibit varying degrees ofunsaturation. In particular embodiments where the edible glycerideincludes an unsaturated fatty acid chain the degree of unsaturation mayrange from one double bond (e.g., oleic acid) up to six double bonds(e.g., docosahexaenoic acid). Further, the fatty acid residues of edibleglycerides suitable for use in the compositions described herein mayform an ester linkage on one (mono-glyceride), two (di-glyceride), orall three glycerol hydroxyl groups (triglyceride). Any hydroxyl groupsnot esterified may be free hydroxyl groups or chemically linked to, forexample, phosphate, inositols, choline, serine, or ethanol amine, suchas found in lecithin.

The edible lipid used in the compositions described herein may also beselected from edible lipids, oils, and fats from plant and animalsources. Plant oils that may be suitable for use in the compositionsdescribed herein include, for example, olive, corn, soy, marine,coconut, palm, palm kernel, cotton seed, peanut, safflower, sesame,sunflower, almond, cashew, macadamia, pecan, pine nut, walnut, lemon,orange, flax seed, and borage oils. Where the edible lipid is a plantoil or is derived from one of the plant oils detailed herein, the ediblelipid may also be selected from cocoa butter or an inter-esterifiedplant oil, such as medium-chain triglycerides formed from one or more ofthe plant oils detailed herein. Animal oils and fats that may besuitable for use in the compositions described herein include fish oilsand dairy derived fats, such as butter.

The nanoparticulate active material can be suspended within the ediblelipid at various concentrations. The compositions described herein canbe prepared with high concentrations of the nanoparticulate material,and in certain embodiments, the nanoparticulate material is suspended inthe edible lipid at a concentration of 200 mg/ml or higher. For example,compositions according to the present description may include thenanoparticulate material suspended within the edible lipid at aconcentration selected from 200 mg/ml or higher, 250 mg/ml or higher,300 mg/ml or higher, 350 mg/ml or higher, 400 mg/ml or higher, 450 mg/mlor higher, and 500 mg/ml or higher.

Compositions according to the present description may additionallyinclude edible colorants, flavorants, and dispersants. A wide variety ofcolorants and flavorants suitable for human and animal consumption arewell known and readily commercially available. Where included in thecompositions described, a dispersant, which may be an edible emulsifier,may serve to help maintain or improve particle size stability within thesuspension and/or bioavailability of the phenolic compound(s) dispersedtherein. The edible dispersant may be selected from, for example,lecithin, and sorbitan fatty acid esters, such as polysorbate 80,steroyl-2-lactate, polyoxyethylene esters, sucrose esters of fattyacids, polyglycerol esters, fatty acid esters of propyleneglycol, andglycerol fatty acid esters. Where included, a dispersant may be providedin a relative amount selected from about 0.01% to about 5% (w/v).

The compositions described herein can be prepared for use in differentcontexts. For example, the compositions may be prepared for oralconsumption as a dietary supplement, a medical food, or even as apharmaceutical formulation. Where prepared as a dietary supplement or amedical food, the phenolic compound(s), edible lipid, and, whereincluded, any colorant, flavorant, or dispersant included in thecomposition can be prepared and provided as a food-grade material suchthat the resulting composition is a food-grade composition suitable foruse as a medical food or dietary supplement. If desired, in otherembodiments, the compositions described herein can be prepared for useas a pharmaceutical composition. In such embodiments, each of thecomponents must be prepared as pharmaceutical grade materials such thata pharmaceutical grade composition is produced.

The compositions described herein may be prepared in any suitable mannerand using any suitable devices. Moreover, compositions according to thepresent description can be prepared for oral consumption or oral orparenteral administration via a variety of delivery devices ormechanisms. For example, the suspension compositions described hereincan be prepared for delivery from any desired metering device, includinga syringe, measuring spoon, cup, or vial, and where desired to easeadministration or delivery, the compositions according to the presentdescription can be metered in pre-measured amounts into syringes,sachets, or capsules, such as gelatin or soft capsules, suited fordelivery of suspension or dispersion compositions. Particularly whenused as a medical food or dietary supplement, the compositions describedherein may also be prepared for distribution over or within other foods,liquids, or dry goods for consumption by the intended subject.

III. Methods

Methods for preparing the compositions described herein are provided. Incertain embodiments, methods for preparing a composition as describedherein include providing a phenolic compound as a nanoparticulatematerial, providing an edible lipid as described herein, and mixing thenanoparticulate phenolic compound material with the edible lipid todisperse the nanoparticulate phenolic compound material and form asuspension composition. In such embodiments, one or more additionalmaterials may also be provided for inclusion in the composition. Forexample, one or more dispersants may be provided and combined with thenanoparticulate phenolic compound material and the edible lipid. Whereone or more dispersants are included, the dispersant may be combinedwith the edible lipid prior to the introduction of the nanoparticulatephenolic compound. Alternatively, one or more dispersants may becombined with the nanoparticulate phenolic compound material prior tocombination of the dispersant and phenolic compound with the ediblelipid. Even further, one or more dispersants may be combined with theedible lipid and nanoparticulate phenolic compound after thenanoparticulate phenolic compound has been dispersed within the ediblelipid.

In yet further embodiments, the compositions described herein can beprepared by forming the nanoparticulate active material in the presenceof the edible lipid, where the edible lipid forms part of the mediumwithin which the size comminution of the phenolic compound takes place.In certain such embodiments, the active material is processed to adesired nanoparticulate PSD using a nanomill, wherein the activematerial is combined with the edible lipid prior to introduction intothe nanomill. In other such embodiments, the active material and ediblelipid are introduced into the nanomill separately (e.g., the phenoliccompound material and edible lipid may be introduced to the millingmedia in separate process steps, the edible lipid may be combined withthe milling media prior to introduction of the active material, or theactive material may be combined with the milling media prior tointroduction of the edible lipid).

In specific embodiments where the active material and edible lipid arecombined in a nanomill, the active material and edible lipid are fedinto the nanomill and milled in a manner that results in a compositioncharacterized by nanoparticulate active material suspended within theedible lipid. In one such embodiment, a suspension of active materialand edible lipid may be fed continuously through the nanomill until asuspension composition containing nanoparticulate active material of adesired PSD is reached. For example, the active material may benanomilled by recirculating a volume of the active material suspendedwithin the edible lipid, followed by one or more single passes throughthe nanomill to reach a composition that includes a phenolic compoundmaterial exhibiting the desired PSD. The particle size of the activematerial suspended within a composition as described herein can becontrolled by adjusting the parameters of the nanomill and the grindingconditions. For example, the particle size produced by nanomilling theactive material or a combination of active material and edible lipid maybe controlled by the size of the milling media, load/suspension weightratio of the milling media, suspension composition (e.g., the amount ofactive material relative to the amount of edible lipid), agitation rate,and milling time.

Depending on the nature of the active material, one or more pre-millingsteps may also be utilized prior to a final nanomilling process.Pre-milling of the active material may be carried out using any suitablemethod and system for size comminution of the active material.Pre-milling may be particularly helpful where the active material, assupplied, exhibits a relatively coarse PSD. By subjecting the activematerial to a pre-milling step, the PSD of the active material can bereduced to better approximate the targeted nanoparticulate PSD and,thereby, reduce the process time required in the final milling process.Such an approach may be particularly advantageous where the activematerial is to be milled in an edible lipid, as the edible lipid and/oractive material may be adversely affected by prolonged exposure to theheat, shear, and grinding forces that could be required to reduce thePSD of a coarse active material to a suitable nanoparticulate range. Apremilling step may also allow sourcing and use of active material thatexhibits relatively coarse initial particle size characteristics, which,in turn, may lead to cost savings in the materials used for preparationof compositions according to the present description.

Methods for administering the physiologically active phenolic compoundsincluded in the compositions described herein are also provided. Theactive materials described herein have several potential nutritional andtherapeutic benefits, with many being recognized as powerfulantioxidants and considered to confer nutritional and therapeuticbenefits to subjects suffering from or at risk of conditions rangingfrom various forms of inflammation to certain forms of cancer.Additionally, physiologically active phenolic compounds disclosed hereinhave been associated with potential cardioprotective and neuroprotectiveeffects, particularly when consumed over time. Even further,physiologically active phenolic compounds specified herein have beenused to ameliorate symptoms associated with changes in hormone levels orproduction in subjects.

The compositions described herein will typically be consumed oradministered orally, and in such embodiments, the amount and frequencywith which the composition is consumed by a subject may depend on thedesired nutritional or therapeutic benefit to be achieved, the nature ofthe phenolic compound(s) included in the composition, and the physicalcharacteristics of the subject (e.g., age, weight, gender). Thecompositions described herein may be administered or consumed, forexample, as a single dose, a regular daily dose, a two-times daily dose,a three-times daily dose, or according to another desired schedule.Moreover, in specific embodiments, a composition as described herein maybe administered to or consumed by a subject in an amount sufficient todeliver a dose of a phenolic compound contained therein that ranges fromabout 50 mg/day to about 10,000 mg/day. In certain such embodiments, thecomposition is delivered to or consumed by the subject in an amountsufficient to deliver a dose of phenolic compound selected from about 50mg/day to about 9,000 mg/day, about 50 mg/day to about 8,000 mg/day,about 50 mg/day to about 2,000 mg/day, about 100 mg/day to about 9,000mg/day, about 100 mg/day to about 5,000 mg/day, about 100 mg/day toabout 4,000 mg/day, about 100 mg/day to about 2,000 mg/day, and about100 mg/day to about 1,000 mg/day.

EXAMPLES Example 1 Preparation of Nano Particulate Compositions ofGenistein

Materials:

Genivida™, a food grade genistein, and Bonistein®, a pharmaceuticalgrade genistein, were obtained from a commercial supplier (DSM,http://www.dsm.com/corporate/home.html) and subjected to particle sizeanalysis. As provided, the Genivida™ material exhibited a D(0.50)particle size 29.5 μm, and the Bonistein® material exhibited a D(0.50)particle size 10.7 μm. FIG. 1 and FIG. 2 illustrate the PSD of theGenivida™ and Bonistein® materials as supplied from DSM.

A food grade, 100% olive oil was obtained for use as an edible lipid.The olive oil was purchased from a supermarket and was a genericMediterranean olive oil blend containing olive oils from Italy, Spain,Tunisia, Turkey, and Morocco.

Methods:

The Genivida™ and Bonistein® materials were dispersed within the oliveoil to provide a suspension composition that included 25% (w/v)genistein. Several samples (≈200 ml per sample) of the 25% w/vgenistein/olive oil suspension were divided and milled using a, MiniCer®nanomill from Netzsch. The samples containing Bonistein® were milled for60 minutes in a 0.3 mm YTZ media (yttria stabilized zirconium oxide(ZrO₂) beads). Samples containing Genivida™ were milled for 60 minutesin a 0.5 mm YTZ media.

A selection of the samples containing Genivida™ were subjected to apre-milling process (referred to or labeled as “pre-grind” samples). Thepre-milling process also utilized a MiniCer® nanomill from Netzsch withYTZ milling media. However, the milling media utilized in thepre-milling step was 1.25 mm YTZ media. Pre-milling was carried out for20 minutes, with the purpose of reducing the D(0.50) particle size ofthe Genivida™ to below 10 μm. After pre-milling, those samples subjectedto the pre-milling procedure were milled in a MiniCer® nanomill for 60minutes in a 0.3 mm YTZ media.

After the milling steps, particle size analysis was completed for eachsample. Milling of the genistein compositions resulted in a sizecomminution of the Bonistein® and Genivida™ materials, with each sampleexhibiting a nanoparticulate PSD (see, FIG. 4). An amount of eachnanosuspension composition was aliquoted into 3.7 cc glass vials, whichwere then stored at ambient temperature.

At approximately two months, samples were taken from each vial andsubjected to laser diffraction particle size analysis and TEM analysis.Particle size analysis and distribution obtained by TEM analysis wasconsistent with that obtained by laser diffraction analysis, and showedthat the genistein material suspended within the samples maintained ananoparticulate PSD.

At the 3-month time point, a sample of each nanosuspension was takenfrom each vial and again subjected to laser diffraction particle sizeanalysis and TEM analysis. Even after three months, the genisteinmaterial suspended in the olive oil remained nano-sized, with theD(0.50) at 3 months in all cases being <300 nm (see, FIGS. 3A-3C).

For each of the samples at each time point, particle size analysisconducted by laser diffraction was carried out using a LA-950 particlesize analyzer from Horiba Scientific, Edison, N.J.

The invention claimed is:
 1. A composition suitable for consumption by asubject, the composition comprising: an edible lipid; and an activematerial suspended within the edible lipid forming a nanoparticulatesuspension, wherein the active material comprises a physiologicallyactive phenolic compound and exhibits a particle size distributioncharacterized by a D (0.50) of 0.5 μm or less.
 2. The compositionaccording to claim 1, wherein the active material exhibits a particlesize distribution characterized by a D (0.50) selected from a D (0.50)of 0.4 μm or less, a D (0.50) of 0.35 μm or less, a D (0.50) of 0.30 μmor less, a D (0.50) of 0.25 μm or less, and a D (0.50) of 0.20 μm orless.
 3. The composition according to claim 2, wherein the activematerial exhibits a particle size distribution characterized by aD(0.50) of 0.3 μm or less.
 4. The composition according to claim 2,wherein the active material exhibits a particle size distributioncharacterized by a D (0.90) of 2.0 μm or less.
 5. The compositionaccording to claim 4, wherein the active material exhibits a particlesize distribution characterized by a D (0.90) selected from a D (0.90)of 1.5 μm or less, a D (0.90) of 1.0 μm or less, and a D (0.90) of 0.5μm or less.
 6. The composition according to claim 2, wherein thephysiologically active phenolic compound is selected from the groupconsisting of isoflavones, curcuminoids, flavonols, stilbenoids, andcombinations thereof.
 7. The composition according to claim 6, whereinthe physiologically active phenolic compound is selected from the groupconsisting of curcumin, desmethoxycurcumin, bis-desmethoxycurcumin,genistein, daidzein, catechin, epicatechin, EGCG, resveratrol, andcombinations of two of more of such compounds.
 8. The compositionaccording to claim 6, wherein the physiologically active phenoliccompound is genistein.
 9. The composition according to claim 6, furthercomprising a dispersant selected from the group consisting of lecithin,polysorbate 80, steroyl-2-lactate, polyoxyethylene esters, sucroseesters of fatty acids, polyglycerol esters, fatty acid esters ofpropylene glycol, and glycerol fatty acid esters.
 10. The compositionaccording to claim 6, wherein the edible lipid is an edible glyceridehaving a chain length ranging from one carbon acetate to 22 carbons, andwherein the edible glyceride is selected from the group consisting ofmono-glycerides, di-glycerides, and triglycerides.
 11. The compositionaccording to claim 6, wherein the edible lipid is an edible plant oil oranimal oil selected from the group consisting of olive, corn, soy,marine, coconut, palm, palm kernel, cotton seed, peanut, safflower,sesame, sunflower, almond, cashew, macadamia, pecan, pine nut, walnut,lemon, orange, flax seed, borage oils, fish oils, and dairy derivedfats.
 12. The composition according to claim 8, wherein the edible lipidcomprises olive oil.
 13. A method for preparing an edible suspension ofa physiologically active phenolic compound, the method comprising:providing an edible lipid; providing an active material comprising aphysiologically active phenolic compound; and milling the activematerial in the presence of the edible lipid to form a nanoparticulatesuspension of the active material suspended within the edible lipid,wherein the active material in the nanoparticulate suspension exhibits aparticle size distribution characterized by a D(0.50) of 0.5 μm or less,and wherein milling the active material in the presence of the ediblelipid comprises nanomilling the active material in a milling media thatcomprises the edible lipid.
 14. The method according to claim 13,wherein, prior to milling the active material in the presence of theedible lipid, the active material is suspended within the lipid andmilling the active material in the presence of the edible lipidcomprises introducing the combined active material and edible lipid intoa nanomill and processing the combined active material and lipid withinthe nanomill until the active material exhibits a desired particle sizedistribution.
 15. The method according to claim 13, wherein milling theactive material in the presence of the edible lipid comprises millingthe active material until the active material exhibits a particle sizedistribution characterized by a D (0.50) selected from a D (0.50) of 0.4μm or less, a D (0.50) of 0.35 μm or less, a D (0.50) of 0.30 μm orless, a D (0.50) of 0.25 μm or less, and a D (0.50) of 0.20 μm or less.16. The method according to claim 15, wherein milling the activematerial in the presence of the edible lipid comprises milling theactive material until the active material exhibits a particle sizedistribution characterized by a D(0.50) of 0.3 μm or less.
 17. Themethod according to claim 15, wherein milling the active material in thepresence of the edible lipid comprises milling the active material untilthe active material exhibits a particle size distribution characterizedby a D (0.90) of 2.0 μm or less.
 18. The method according to claim 17,wherein milling the active material in the presence of the edible lipidcomprises milling the active material until the active material exhibitsa particle size distribution characterized by a D (0.90) selected from aD (0.90) of 1.5 μm or less, a D (0.90) of 1.0 μm or less, and a D (0.90)of 0.5 μm or less.
 19. The method according to claim 15, wherein thephysiologically active phenolic compound is selected from the groupconsisting of isoflavones, curcuminoids, flavonols, stilbenoids, andcombinations thereof.
 20. The method according to claim 19, wherein thephysiologically active phenolic compound is selected from the groupconsisting of curcumin, desmethoxycurcumin, bis-desmethoxycurcumin,genistein, daidzein, catechin, epicatechin, EGCG, resveratrol, andcombinations of two of more of such compounds.
 21. The method accordingto claim 20, wherein the physiologically active phenolic compound isgenistein.
 22. The method according to claim 19, further comprisingproviding a dispersant, wherein the dispersant is blended with theedible lipid and the active material, and wherein the dispersant isselected from the group consisting of lecithin, polysorbate 80,steroyl-2-lactate, polyoxyethylene esters, sucrose esters of fattyacids, polyglycerol esters, fatty acid esters of propyleneglycol, andglycerol fatty acid esters.
 23. The method according to claim 19,wherein the edible lipid is an edible glyceride having a chain lengthranging from one carbon acetate to 22 carbons, and wherein the edibleglyceride is selected from the group consisting of mono-glycerides,di-glycerides, and triglycerides.
 24. The method according to claim 19,wherein the edible lipid is an edible plant oil or animal oil selectedfrom the group consisting of olive, corn, soy, marine, coconut, palm,palm kernel, cotton seed, peanut, safflower, sesame, sunflower, almond,cashew, macadamia, pecan, pine nut, walnut, lemon, orange, flax seed,borage oils, fish oils, and dairy derived fats.
 25. The compositionaccording to claim 1, wherein the concentration of the active materialin the nanoparticulate suspension is 200 mg/ml or higher.
 26. Thecomposition according to claim 25, wherein the concentration of theactive material in the nanoparticulate suspension is selected from aconcentration of 250 mg/ml or higher, a concentration of 300 mg/ml orhigher, a concentration of 350 mg/ml or higher, a concentration of 400mg/ml or higher, a concentration of 450 mg/ml or higher, and aconcentration of 500 mg/ml or higher.
 27. The method according to claim13, wherein the concentration of the active material in thenanoparticulate suspension is 200 mg/ml or higher.
 28. The methodaccording to claim 27, wherein the concentration of the active materialin the nanoparticulate suspension is selected from a concentration of250 mg/ml or higher, a concentration of 300 mg/ml or higher, aconcentration of 350 mg/ml or higher, a concentration of 400 mg/ml orhigher, a concentration of 450 mg/ml or higher, and a concentration of500 mg/ml or higher.
 29. The method according to claim 19, wherein theconcentration of the active material in the nanoparticulate suspensionis 200 mg/ml or higher.
 30. The method according to claim 29, whereinthe concentration of the active material in the nanoparticulatesuspension is selected from a concentration of 250 mg/ml or higher, aconcentration of 300 mg/ml or higher, a concentration of 350 mg/ml orhigher, a concentration of 400 mg/ml or higher, a concentration of 450mg/ml or higher, and a concentration of 500 mg/ml or higher.